SummaryRMgm-60
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene disruption |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 15970588 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. berghei |
Parent strain/line | P. berghei ANKA |
Name parent line/clone | P. berghei ANKA 2.34 |
Other information parent line | P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943). |
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The mutant parasite was generated by | |
Name PI/Researcher | L. Reininger, C. Doerig |
Name Group/Department | INSERM U609 |
Name Institute | Wellcome Centre for Molecular Parasitology, University of Glasgow |
City | Glasgow |
Country | Scotland, UK |
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Name of the mutant parasite | |
RMgm number | RMgm-60 |
Principal name | 37.7; 37.9 |
Alternative name | pbnek-4-KO |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Not different from wild type |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Gamete formation is not affected. Macrogametes become activated but fail to develop into ookinetes. Fertilisation occurs, but development is arrested at 2N zygote stage. The defect is female gamete specific; male gametes are fertile and able to fertilize wild-type female gametes. |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype |
Protein (function) Phenotype Additional information Other mutants |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0616700 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0719200 | ||||||||||||||||||||||||
Gene product | NIMA related kinase 4 | ||||||||||||||||||||||||
Gene product: Alternative name | serine/threonine protein kinase, Pfnek-4 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | Plasmid double cross-over | ||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Partial | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | To disrupt pbnek-4, most of the kinase domain (residues 84-247) was replaced with a pyrimethamine-resistant allele of the dhfr/ts gene from T. gondii | ||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | tgdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | The pbnek-4-KO line has been complemented with an intact copy of the pbnek4 gene, tagged with a carboxy-terminal c-myc epitope tag (insertion vector). Complementation succeeded in restoring ookinete formation, although not to the level of the wild type. | ||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
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